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Application of recombinant immunoregulatory protein of ganoderma lucidum in preparation of drug for treating focal cerebral ischemia

a technology of immunoregulatory protein and ganoderma lucidum, which is applied in the field of biological pharmacy, can solve the problems of serious threat to the health and life of people, and achieve the effect of significant therapeutic effect on the focal cerebral ischemia

Inactive Publication Date: 2016-07-14
ZHANG XITIAN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention describes a method for creating models of cerebral ischemia in rats using a modified version of the Zea Longa method. The rats were divided into a negative control group, a positive drug control group, and three experimental groups, each containing 7, 15, or 70 μg·kg−1 of rLZ-8. The rLZ-8 was injected intraperitoneally twice a day for 7 days. The animals were tested for their neurological function using a grading system, and the results showed a significant improvement in the rLZ-8 high-dosage group compared to the model group. Hematoxylin-Eosin staining showed a protective effect on brain tissue. The high-dosage group also showed a decrease in inflammatory response and a lower number of apoptotic cells. The invention provides a reliable and effective method for studying cerebral ischemia in rats, which can help in the development of new therapies for this condition.

Problems solved by technology

The cerebrovascular accident, also called cerebral apoplexy and stroke, is the common disease and the frequently-occurring disease of the middle aged and elderly people, which seriously threatens the health and the life of people.

Method used

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  • Application of recombinant immunoregulatory protein of ganoderma lucidum in preparation of drug for treating focal cerebral ischemia
  • Application of recombinant immunoregulatory protein of ganoderma lucidum in preparation of drug for treating focal cerebral ischemia
  • Application of recombinant immunoregulatory protein of ganoderma lucidum in preparation of drug for treating focal cerebral ischemia

Examples

Experimental program
Comparison scheme
Effect test

first example

Therapeutic Effect of rLZ-8 on Neurological Function Injury

[0016]1. Experimental Material

[0017]Rats, having a weight of 250-300 g, half males and half females, were provided by Laboratory Animal Center of Jilin University. GM-1 was provided by Beijing Four-ring Pharmaceutical Co., Ltd. Surgical instruments.

[0018]2. Experimental Method

[0019]Preparing rat focal cerebral ischemia models through a modified Zea Longa method; grading the rats which were fully conscious after operation according to a modified NSS grading standard (16-point system); arranging a sham operation group, a model group, a positive drug control group, a rLZ-8 high-dosage group (70 μg·kg−1), a rLZ-8 middle-dosage group (35 μg·kg−1) and a rLZ-8 low-dosage group (17.5 μg·kg−1); and injecting intraperitoneally, twice a day, wherein the same volume of physiological saline was injected into the sham operation group and the model group, for consecutive 7 days. A positive drug in the positive drug control group was the GM...

second example

Influence of rLZ-8 on Weights of Rats

[0022]1. Experimental Method

[0023]Weighing rats before modeling; preparing rat focal cerebral ischemia models through a modified Zea Longa method; weighing the rats on the first postoperative day; and weighing the rats after consecutively administrating a drug for 7 days.

[0024]2. Experimental Results

[0025]As shown in Table 2 and FIG. 1, before modeling, weights of the rats of each group had no significant difference; after modeling, the weights of the rats of each group were significantly decreased; and after consecutively administrating the drug for 7 days, the weights of the rats of the rLZ-8 high-dosage group were increased, illustrating that the rLZ-8 is able to reduce an injury caused by cerebral ischemia.

TABLE 2Changes of weights of rats of each group (x± s)0th day after1st day after8th day afterGroupmodelingmodelingmodelingSham268.33 ± 13.87267.17 ± 13.73295.00 ± 24.22 M261.50 ± 16.57 235.67 ± 18.97** 230.17 ± 26.49**GM-1266.17 ± 21.21246....

third example

HE Staining Experiment

[0026]1. Experimental Method

[0027]Perfusing and fixing: anesthetizing a rat by chloral hydrate and fixing the rat in a supine position; processing the rat with thoracotomy to expose a heart; inserting a perfusing needle into an aorta through an apex of a left ventricle; firstly perfusing the rat quickly by physiological saline, and meanwhile, cutting off a right auricle and clamping the aorta by a hemostatic forceps; when no blood was in an effluent liquid, quickly perfusing the rat by fixative, and stopping perfusing when a body of the rat became stiff; cutting off a head to remove out a brain tissue; preparing a coronary brain section through a rat brain section mold; fixing the coronary brain section by formalin; selecting a brain section (the third brain section) which was horizontal with an optic chiasma; embedding the brain section which was horizontal with an optic chiasma by conventional paraffin and sectioning.

[0028]HE staining: (1) dewaxing the embedd...

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Abstract

An application of a recombinant immunoregulatory protein of ganoderma lucidum (rLZ-8) expressed by Pichia pastoris in a preparation of a drug for treating focal cerebral ischemia is provided. The rLZ-8 is able to treat a neurological function injury, decrease a neurological severity score caused by a cerebral injury and decrease an ED-1 positive cell number, so as to decrease an inflammatory response and inhibit apoptosis.

Description

CROSS REFERENCE OF RELATED APPLICATION[0001]This is a U.S. National Stage under 35 U.S.C 371 of the International Application PCT / CN2014 / 079816, filed Jun. 13, 2014, which claims priority under 35 U.S.C. 119(a-d) to CN 201310402259.4, filed Sep. 6, 2013.BACKGROUND OF THE PRESENT INVENTION[0002]1. Field of Invention[0003]The present invention belongs to a field of biological pharmacy, and relates to an application of a recombinant immunoregulatory protein of ganoderma lucidum (rLZ-8) expressed by Pichia pastoris in a preparation of a drug for treating focal cerebral ischemia.[0004]2. Description of Related Arts[0005]The ganoderma lucidum is a precious medicinal fungus of the traditional Chinese pharmacy, belonging to ganodermataceae of polyporales of basidiomycetes. The immunoregulatory protein of ganoderma lucidum is a fungal immunoregulatory protein which is separated and purified from a ganoderma lucidum fruit body extract firstly by Kino et al. The immunoregulatory protein of gan...

Claims

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Application Information

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IPC IPC(8): A61K38/16
CPCA61K38/168A61P25/00A61P29/00A61P9/10
Inventor ZHANG, XITIANSUN, FEILIANG, CHONGYANG
Owner ZHANG XITIAN
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